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2 ATP + L-valine + 2 L-Baa = 2 ADP + 2 phosphate + L-Val-(L-Baa)2
3 ATP + 3 L-valine + L-Zaa = 3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-amino-acid-L-Zaa
ATP + 2 L-valine + L-Yaa = 2 ADP + 2 phosphate + L-Val-L-Val-L-Yaa
n-1 ATP + n L-Xaa = n-1 ADP + n-1 phosphate + (Xaa)n
2 ATP + L-valine + 2 L-Baa = 2 ADP + 2 phosphate + L-Val-(L-Baa)2

Baa: Leu, Ile, Met
2 ATP + L-valine + 2 L-Baa = 2 ADP + 2 phosphate + L-Val-(L-Baa)2
Baa: Leu, Ile, Met
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2 ATP + L-valine + 2 L-Baa = 2 ADP + 2 phosphate + L-Val-(L-Baa)2
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3 ATP + 3 L-valine + L-Zaa = 3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-amino-acid-L-Zaa

Zaa: Arg, His, Gln, Asn or Phe
3 ATP + 3 L-valine + L-Zaa = 3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-amino-acid-L-Zaa
Zaa: Arg, His, Gln, Asn or Phe
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3 ATP + 3 L-valine + L-Zaa = 3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-amino-acid-L-Zaa
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ATP + 2 L-valine + L-Yaa = 2 ADP + 2 phosphate + L-Val-L-Val-L-Yaa

Yaa: Arg, Lys, His, Gln, Asn, Ala, Ser, Thr, Gly, Leu, Ile, Met, Phe or Trp
ATP + 2 L-valine + L-Yaa = 2 ADP + 2 phosphate + L-Val-L-Val-L-Yaa
Yaa: Arg, Lys, His, Gln, Asn, Ala, Ser, Thr, Gly, Leu, Ile, Met, Phe or Trp
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ATP + 2 L-valine + L-Yaa = 2 ADP + 2 phosphate + L-Val-L-Val-L-Yaa
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n-1 ATP + n L-Xaa = n-1 ADP + n-1 phosphate + (Xaa)n

Xaa: Val, Leu, or Met. n = 2-5
n-1 ATP + n L-Xaa = n-1 ADP + n-1 phosphate + (Xaa)n
the enzyme catalyzes the synthesis of oligopeptides from unprotected L-amino acids in an ATP-dependent manner, showing synthesis of various oligopeptides consisting of two to five amino acids, e.g. synthesis of Val-Val-Val tripeptide by the enzyme RizB. The enzyme shows highest activity with Val, Ile, Leu, and Met as substrates. RizB shows specificity toward N-terminal substrates, but is relaxed in the specificity for the C-terminus
n-1 ATP + n L-Xaa = n-1 ADP + n-1 phosphate + (Xaa)n
Xaa: Val, Leu, or Met. n = 2-5
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n-1 ATP + n L-Xaa = n-1 ADP + n-1 phosphate + (Xaa)n
the enzyme catalyzes the synthesis of oligopeptides from unprotected L-amino acids in an ATP-dependent manner, showing synthesis of various oligopeptides consisting of two to five amino acids, e.g. synthesis of Val-Val-Val tripeptide by the enzyme RizB. The enzyme shows highest activity with Val, Ile, Leu, and Met as substrates. RizB shows specificity toward N-terminal substrates, but is relaxed in the specificity for the C-terminus
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n-1 ATP + n L-Xaa = n-1 ADP + n-1 phosphate + (Xaa)n
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2 ATP + 2 L-Val + Gly
2 ADP + 2 phosphate + L-Val-L-Val-Gly
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2 ATP + 2 L-Val + L-Ala
2 ADP + 2 phosphate + L-Val-L-Val-L-Ala
2 ATP + 2 L-Val + L-Arg
2 ADP + 2 phosphate + L-Val-L-Val-L-Arg
2 ATP + 2 L-Val + L-Asn
2 ADP + 2 phosphate + L-Val-L-Val-L-Asn
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2 ATP + 2 L-Val + L-Gln
2 ADP + 2 phosphate + L-Val-L-Val-L-Gln
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2 ATP + 2 L-Val + L-His
2 ADP + 2 phosphate + L-Val-L-Val-L-His
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2 ATP + 2 L-Val + L-Ile
2 ADP + 2 phosphate + L-Val-L-Val-L-Ile
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2 ATP + 2 L-Val + L-Leu
2 ADP + 2 phosphate + L-Val-L-Val-L-Leu
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2 ATP + 2 L-Val + L-Lys
2 ADP + 2 phosphate + L-Val-L-Val-L-Lys
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2 ATP + 2 L-Val + L-Met
2 ADP + 2 phosphate + L-Val-L-Val-L-Met
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2 ATP + 2 L-Val + L-Phe
2 ADP + 2 phosphate + L-Val-L-Val-L-Phe
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2 ATP + 2 L-Val + L-Ser
2 ADP + 2 phosphate + L-Val-L-Val-L-Ser
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2 ATP + 2 L-Val + L-Thr
2 ADP + 2 phosphate + L-Val-L-Val-L-Thr
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2 ATP + 2 L-Val + L-Trp
2 ADP + 2 phosphate + L-Val-L-Val-L-Trp
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2 ATP + 3 L-leucine
2 ADP + 2 phosphate + L-Leu-L-Leu-L-Leu
the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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2 ATP + 3 L-methionine
2 ADP + 2 phosphate + L-Met-L-Met-L-Met
the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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2 ATP + 3 L-valine
2 ADP + 2 phosphate + L-Val-L-Val-L-Val
2 ATP + L-Val + 2 L-Leu
2 ADP + 2 phosphate + L-Val-L-Leu-L-Leu
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2 ATP + L-Val + 2 L-Met
2 ADP + 2 phosphate + L-Met-L-Met-L-Met
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2 ATP + L-Val + L-Arg-L-Ser
2 ADP + phosphate + L-Val-L-Arg-L-Ser
no homopeptides of Val-Val and Val-Val-Val are detected. This suggests that the enzyme preferentially uses dipeptides as the C-terminal substrate
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2 ATP + L-Val + L-Val + L-Arg-L-Ser
2 ADP + 2 phosphate + L-Val-L-Val-L-Arg-L-Ser
no homopeptides of Val-Val and Val-Val-Val are detected. This suggests that the enzyme preferentially uses dipeptides as the C-terminal substrate
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2 ATP + Val + 2 L-Ile
2 ADP + 2 phosphate + L-Val-L-Ile-L-Ile
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3 ATP + 3 L-Val + L-Arg
3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-Arg
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3 ATP + 3 L-Val + L-Gln
3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-Gln
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3 ATP + 3 L-Val + L-His
3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-His
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3 ATP + 3 L-Val + L-Phe
3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-Phe
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3 ATP + 4 L-leucine
3 ADP + 3 phosphate + L-Leu-L-Leu-L-Leu-L-Leu
the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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3 ATP + 4 L-methionine
3 ADP + 3 phosphate + L-Met-L-Met-L-Met-L-Met
the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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3 ATP + 4 L-valine
3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-Val
4 ATP + 5 L-leucine
4 ADP + 4 phosphate + L-Leu-L-Leu-L-Leu-L-Leu-L-Leu
the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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4 ATP + 5 L-methionine
4 ADP + 4 phosphate + L-Met-L-Met-L-Met-L-Met-L-Met
the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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4 ATP + 5 L-valine
4 ADP + 4 phosphate + L-Val-L-Val-L-Val-L-Val-L-Val
the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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ATP + 2 L-leucine
ADP + phosphate + L-Leu-L-Leu
the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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ATP + 2 L-methionine
ADP + phosphate + L-Met-L-Met
the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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ATP + 2 L-valine
ADP + phosphate + L-Val-L-Val
ATP + L-Val + L-Arg
ADP + phosphate + L-Val-L-Arg
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ATP + L-Val + L-Arg-L-Ser
ADP + phosphate + L-Val-L-Arg-L-Ser + L-Val-L-Val-L-Arg-L-Ser
no homopeptides of Val-Val and Val-Val-Val are detected. The enzyme preferentially uses dipeptides as the C-terminal substrates
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ATP + L-Val + L-Arg-NHOH
ADP + phosphate + L-Val-L-Arg-NHOH + L-Val-L-Val-L-Arg-NHOH
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ATP + L-Val + L-Phe-NHOH
ADP + phosphate + L-Val-L-Phe-NHOH + L-Val-L-Val-L-Phe-NHOH
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ATP + L-valine + L-Val-L-Val
ADP + phosphate + L-Val-L-Val-L-Val
ATP + L-valine + L-Val-L-Val-L-Val
ADP + phosphate + L-Val-L-Val-L-Val-L-Val
additional information
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2 ATP + 2 L-Val + L-Ala

2 ADP + 2 phosphate + L-Val-L-Val-L-Ala
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2 ATP + 2 L-Val + L-Ala
2 ADP + 2 phosphate + L-Val-L-Val-L-Ala
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2 ATP + 2 L-Val + L-Arg

2 ADP + 2 phosphate + L-Val-L-Val-L-Arg
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2 ATP + 2 L-Val + L-Arg
2 ADP + 2 phosphate + L-Val-L-Val-L-Arg
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2 ATP + 3 L-valine

2 ADP + 2 phosphate + L-Val-L-Val-L-Val
the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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2 ATP + 3 L-valine
2 ADP + 2 phosphate + L-Val-L-Val-L-Val
the enzyme synthesizes homo-oligomers of two to four amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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2 ATP + 3 L-valine
2 ADP + 2 phosphate + L-Val-L-Val-L-Val
the enzyme synthesizes homo-oligomers of two to four amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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3 ATP + 4 L-valine

3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-Val
the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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3 ATP + 4 L-valine
3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-Val
the enzyme synthesizes homo-oligomers of two to four amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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3 ATP + 4 L-valine
3 ADP + 3 phosphate + L-Val-L-Val-L-Val-L-Val
the enzyme synthesizes homo-oligomers of two to four amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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ATP + 2 L-valine

ADP + phosphate + L-Val-L-Val
the enzyme synthesizes homo-oligomers of two to five amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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ATP + 2 L-valine
ADP + phosphate + L-Val-L-Val
the enzyme synthesizes homo-oligomers of two to four amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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ATP + 2 L-valine
ADP + phosphate + L-Val-L-Val
the enzyme synthesizes homo-oligomers of two to four amino acids. ATP cannot by replaced with GTP, CTP or dTTP
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ATP + L-valine + L-Val-L-Val

ADP + phosphate + L-Val-L-Val-L-Val
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ATP + L-valine + L-Val-L-Val
ADP + phosphate + L-Val-L-Val-L-Val
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ATP + L-valine + L-Val-L-Val-L-Val

ADP + phosphate + L-Val-L-Val-L-Val-L-Val
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ATP + L-valine + L-Val-L-Val-L-Val
ADP + phosphate + L-Val-L-Val-L-Val-L-Val
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additional information

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dipeptides, tripeptides, tetrapeptides and pentapeptides are synthesized in the reaction mixtures containing Val, Leu, or Met as substrate. When Ile is used, the longest peptides are tetrapeptides. No peptides are detected when the other 15 amino acids are used as substrates. When Leu, Ile, or Met is used as substrate with Val, heteropeptides containing two residues of Leu, Ile, and Met are detected. No heteropeptides containing Tyr. Pro, Cys, Asp or Glu are detected. When other amino acids that are not usable for homopeptide synthesis are used as substrate, heteropeptides containing only one amino acid residue of these amino acids are detected. Substrate recognition at the N-terminus influences the resulting heteropeptide sequence. The enzyme does not participate in peptide-to-peptide ligation. Very low activity with D-amino acids
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additional information
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the enzyme catalyzes the synthesis of oligopeptides from unprotected L-amino acids in an ATP-dependent manner. Synthesis of various oligopeptides consisting of two to five amino acids, e.g. synthesis of Val-Val-Val tripeptide by the enzyme RizB. The enzyme shows highest activity with Val, Ile, Leu, and Met as substrates. In the reaction with Val, tetramers are the longest products synthesized, trimer are formed with Co2+. RizB shows specificity toward N-terminal substrates, but is relaxed in the specificity for the C-terminus, mass spectrometric reaction analysis, overview
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additional information
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dipeptides, tripeptides, tetrapeptides and pentapeptides are synthesized in the reaction mixtures containing Val, Leu, or Met as substrate. When Ile is used, the longest peptides are tetrapeptides. No peptides are detected when the other 15 amino acids are used as substrates. When Leu, Ile, or Met is used as substrate with Val, heteropeptides containing two residues of Leu, Ile, and Met are detected. No heteropeptides containing Tyr. Pro, Cys, Asp or Glu are detected. When other amino acids that are not usable for homopeptide synthesis are used as substrate, heteropeptides containing only one amino acid residue of these amino acids are detected. Substrate recognition at the N-terminus influences the resulting heteropeptide sequence. The enzyme does not participate in peptide-to-peptide ligation. Very low activity with D-amino acids
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additional information
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the enzyme catalyzes the synthesis of oligopeptides from unprotected L-amino acids in an ATP-dependent manner. Synthesis of various oligopeptides consisting of two to five amino acids, e.g. synthesis of Val-Val-Val tripeptide by the enzyme RizB. The enzyme shows highest activity with Val, Ile, Leu, and Met as substrates. In the reaction with Val, tetramers are the longest products synthesized, trimer are formed with Co2+. RizB shows specificity toward N-terminal substrates, but is relaxed in the specificity for the C-terminus, mass spectrometric reaction analysis, overview
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